LOCUS Exported 8045 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Gateway ORF clone of human TSC2 [NM_001114382.1] without stop codon (for C-terminal fusions). ACCESSION . VERSION . KEYWORDS R777-E356 Hs.TSC2-nostop SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8045) TITLE Esposito lab REFERENCE 2 (bases 1 to 8045) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8045 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 23..45 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 37..54 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 38..54 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 84..183 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(5536..5635) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 6012..6803 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 6896..7484 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7385..7404 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7638..7655 /label=L4440 /note="L4440 vector, forward primer" terminator 7814..7841 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 7933..8019 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" ORIGIN 1 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaaa 61 attcgagctc ggtccgagga tccgaaataa tgattttatt ttgactgata gtgacctgtt 121 cgttgcaaca aattgataag caatgctttt ttataatgcc aactttgtac aaaaaagttg 181 gcaccatggc caaaccaaca agcaaagatt caggcttgaa ggagaagttt aagattctgt 241 tgggactggg aacaccgagg ccaaatccca ggtctgcaga gggtaaacag acggagttta 301 tcatcaccgc ggaaatactg agagaactga gcatggaatg tggcctcaac aatcgcatcc 361 ggatgatagg gcagatttgt gaagtcgcaa aaaccaagaa atttgaagag cacgcagtgg 421 aagcactctg gaaggcggtc gcggatctgt tgcagccgga gcggccgctg gaggcccggc 481 acgcggtgct ggctctgctg aaggccatcg tgcaggggca gggcgagcgt ttgggggtcc 541 tcagagccct cttctttaag gtcatcaagg attacccttc caacgaagac cttcacgaaa 601 ggctggaggt tttcaaggcc ctcacagaca atgggagaca catcacctac ttggaggaag 661 agctggctga ctttgtcctg cagtggatgg atgttggctt gtcctcggaa ttccttctgg 721 tgctggtgaa cttggtcaaa ttcaatagct gttacctcga cgagtacatc gcaaggatgg 781 ttcagatgat ctgtctgctg tgcgtccgga ccgcgtcctc tgtggacata gaggtctccc 841 tgcaggtgct ggacgccgtg gtctgctaca actgcctgcc ggctgagagc ctcccgctgt 901 tcatcgttac cctctgtcgc accatcaacg tcaaggagct ctgcgagcct tgctggaagc 961 tgatgcggaa cctccttggc acccacctgg gccacagcgc catctacaac atgtgccacc 1021 tcatggagga cagagcctac atggaggacg cgcccctgct gagaggagcc gtgttttttg 1081 tgggcatggc tctctgggga gcccaccggc tctattctct caggaactcg ccgacatctg 1141 tgttgccatc attttaccag gccatggcat gtccgaacga ggtggtgtcc tatgagatcg 1201 tcctgtccat caccaggctc atcaagaagt ataggaagga gctccaggtg gtggcgtggg 1261 acattctgct gaacatcatc gaacggctcc ttcagcagct ccagaccttg gacagcccgg 1321 agctcaggac catcgtccat gacctgttga ccacggtgga ggagctgtgt gaccagaacg 1381 agttccacgg gtctcaggag agatactttg aactggtgga gagatgtgcg gaccagaggc 1441 ctgagtcctc cctcctgaac ctgatctcct atagagcgca gtccatccac ccggccaagg 1501 acggctggat tcagaacctg caggcgctga tggagagatt cttcaggagc gagtcccgag 1561 gcgccgtgcg catcaaggtg ctggacgtgc tgtcctttgt gctgctcatc aacaggcagt 1621 tctatgagga ggagctgatt aactcagtgg tcatctcgca gctctcccac atccccgagg 1681 ataaagacca ccaggtccga aagctggcca cccagttgct ggtggacctg gcagagggct 1741 gccacacaca ccacttcaac agcctgctgg acatcatcga gaaggtgatg gcccgctccc 1801 tctccccacc cccggagctg gaagaaaggg atgtggccgc atactcggcc tccttggagg 1861 atgtgaagac agccgtcctg gggcttctgg tcatccttca gaccaagctg tacaccctgc 1921 ctgcaagcca cgccacgcgt gtgtatgaga tgctggtcag ccacattcag ctccactaca 1981 agcacagcta caccctgcca atcgcgagca gcatccggct gcaggccttt gacttcctgt 2041 tgctgctgcg ggccgactca ctgcaccgcc tgggcctgcc caacaaggat ggagtcgtgc 2101 ggttcagccc ctactgcgtc tgcgactaca tggagccaga gagaggctct gagaagaaga 2161 ccagcggccc cctttctcct cccacagggc ctcctggccc ggcgcctgca ggccccgccg 2221 tgcggctggg gtccgtgccc tactccctgc tcttccgcgt cctgctgcag tgcttgaagc 2281 aggagtctga ctggaaggtg ctgaagctgg ttctgggcag gctgcctgag tccctgcgct 2341 ataaagtgct catctttact tccccttgca gtgtggacca gctgtgctct gctctctgct 2401 ccatgctttc aggcccaaag acactggagc ggctccgagg cgccccagaa ggcttctcca 2461 gaactgactt gcacctggcc gtggttccag tgctgacagc attaatctct taccataact 2521 acctggacaa aaccaaacag cgcgagatgg tctactgcct ggagcagggc ctcatccacc 2581 gctgtgccag ccagtgcgtc gtggccttgt ccatctgcag cgtggagatg cctgacatca 2641 tcatcaaggc gctgcctgtt ctggtggtga agctcacgca catctcagcc acagccagca 2701 tggccgtccc actgctggag ttcctgtcca ctctggccag gctgccgcac ctctacagga 2761 actttgccgc ggagcagtat gccagtgtgt tcgccatctc cctgccgtac accaacccct 2821 ccaagtttaa tcagtacatc gtgtgtctgg cccatcacgt catagccatg tggttcatca 2881 ggtgccgcct gcccttccgg aaggattttg tccctttcat cactaagggc ctgcggtcca 2941 atgtcctctt gtcttttgat gacacccccg agaaggacag cttcagggcc cggagtacta 3001 gtctcaacga gagacccaag agtctgagga tagccagacc ccccaaacaa ggcttgaata 3061 actctccacc cgtgaaagaa ttcaaggaga gctctgcagc cgaggccttc cggtgccgca 3121 gcatcagtgt gtctgaacat gtggtccgca gcaggataca gacgtccctc accagtgcca 3181 gcttggggtc tgcagatgag aactccgtgg cccaggctga cgatagcctg aaaaacctcc 3241 acctggagct cacggaaacc tgtctggaca tgatggctcg atacgtcttc tccaacttca 3301 cggctgtccc gaagaggtct cctgtgggcg agttcctcct agcgggtggc aggaccaaaa 3361 cctggctggt tgggaacaag cttgtcactg tgacgacaag cgtgggaacc gggacccggt 3421 cgttactagg cctggactcg ggggagctgc agtccggccc ggagtcgagc tccagccccg 3481 gggtgcatgt gagacagacc aaggaggcgc cggccaagct ggagtcccag gctgggcagc 3541 aggtgtcccg tggggcccgg gatcgggtcc gttccatgtc ggggggccat ggtcttcgag 3601 ttggcgccct ggacgtgccg gcctcccagt tcctgggcag tgccacttct ccaggaccac 3661 ggactgcacc agccgcgaaa cctgagaagg cctcagctgg cacccgggtt cctgtgcagg 3721 agaagacgaa cctggcggcc tatgtgcccc tgctgaccca gggctgggcg gagatcctgg 3781 tccggaggcc cacagggaac accagctggc tgatgagcct ggagaacccg ctcagccctt 3841 tctcctcgga catcaacaac atgcccctgc aggagctgtc taacgccctc atggcggctg 3901 agcgcttcaa ggagcaccgg gacacagccc tgtacaagtc actgtcggtg ccggcagcca 3961 gcacggccaa accccctcct ctgcctcgct ccaacacaga ctccgccgtg gtcatggagg 4021 agggaagtcc gggcgaggtt cctgtgctgg tggagccccc agggttggag gacgttgagg 4081 cagcgctagg catggacagg cgcacggatg cctacagcag gtcgtcctca gtctccagcc 4141 aggaggagaa gtcgctccac gcggaggagc tggttggcag gggcatcccc atcgagcgag 4201 tcgtctcctc ggagggtggc cggccctctg tggacctctc cttccagccc tcgcagcccc 4261 tgagcaagtc cagctcctct cccgagctgc agactctgca ggacatcctc ggggaccctg 4321 gggacaaggc cgacgtgggc cggctgagcc ctgaggttaa ggcccggtca cagtcaggga 4381 ccctggacgg ggaaagtgct gcctggtcgg cctcgggcga agacagtcgg ggccagcccg 4441 agggtccctt gccttccagc tccccccgct cgcccagtgg cctccggccc cgaggttaca 4501 ccatctccga ctcggcccca tcacgcaggg gcaagagagt agagagggac gccttaaaga 4561 gcagagccac agcctccaat gcagagaaag tgccaggcat caaccccagt ttcgtgttcc 4621 tgcagctcta ccattccccc ttctttggcg acgagtcaaa caagccaatc ctgctgccca 4681 atgagtcaca gtcctttgag cggtcggtgc agctcctcga ccagatccca tcatacgaca 4741 cccacaagat cgccgtcctg tatgttggag aaggccagag caacagcgag ctcgccatcc 4801 tgtccaatga gcatggctcc tacaggtaca cggagttcct gacgggcctg ggccggctca 4861 tcgagctgaa ggactgccag ccggacaagg tgtacctggg aggcctggac gtgtgtggtg 4921 aggacggcca gttcacctac tgctggcacg atgacatcat gcaagccgtc ttccacatcg 4981 ccaccctgat gcccaccaag gacgtggaca agcaccgctg cgacaagaag cgccacctgg 5041 gcaacgactt tgtgtccatt gtctacaatg actccggtga ggacttcaag cttggcacca 5101 tcaagggcca gttcaacttt gtccacgtga tcgtcacccc gctggactac gagtgcaacc 5161 tggtgtccct gcagtgcagg aaagacatgg agggccttgt ggacaccagc gtggccaaga 5221 tcgtgtctga ccgcaacctg cccttcgtgg cccgccagat ggccctgcac gcaaatatgg 5281 cctcacaggt gcatcatagc cgctccaacc ccaccgatat ctacccctcc aagtggattg 5341 cccggctccg ccacatcaag cggctccgcc agcggatctg cgaggaagcc gcctactcca 5401 accccagcct acctctggtg caccctccgt cccatagcaa agcccctgca cagactccag 5461 ccgagcccac acctggctat gaggtgggcc agcggaagcg cctcatctcc tcggtggagg 5521 acttcaccga gtttgtgtca actttcttgt acaaagttgg cattatagga aagcattgct 5581 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgcatgc 5641 aagcttagat cctctagacc agccaggaca gaaatgcctc gacttcgctg ctacccaagg 5701 ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct 5761 gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct 5821 tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt 5881 tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc 5941 gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa 6001 agttaaacat tatgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag 6061 ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg 6121 cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa 6181 ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc 6241 ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca 6301 ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca 6361 ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa 6421 aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg 6481 ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc 6541 ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg 6601 cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc 6661 cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag 6721 atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga 6781 tcaccaaggt agtcggcaaa taaccctcga gccacccatg accaaaatcc cttaacgtga 6841 gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag 6901 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg 6961 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca 7021 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga 7081 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca 7141 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc 7201 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca 7261 ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa 7321 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc 7381 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc 7441 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg 7501 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat 7561 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca 7621 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca 7681 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg 7741 actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc caggaagagt 7801 ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc 7861 agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc 7921 cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg 7981 aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag ttccctactc 8041 tcgcg //